Technical Field
[0001] The present invention relates to a formulation for treating Hunter syndrome and a
method for treating Hunter syndrome with the same.
Background Art
[0002] Hunter syndrome or mucopolysaccharidosis type II is one of the lysosomal storage
diseases (LSD) in which mucopolysaccharides such as glycosaminoglycan (GAG) do not
decompose to thereby accumulate in lysosomes due to a deficiency of iduronate-2-sulfatase
(IDS). GAG accumulates in all cells of the body and causes various symptoms, which
include prominent facial features, large head, and abdominal distension due to hypertrophy
of the liver or spleen, and are accompanied by hearing loss, heart valve diseases,
obstructive respiratory diseases, and sleep apnea. It may also involve a limitation
of joint motion as well as nervous system symptoms and developmental delay caused
by invasion of the central nervous system. Hunter syndrome is known to occur in about
1 out of 162,000 people and is inherited as an X-linked recessive form, which causes
great pains for not only the patients but also their family members.
[0003] Up to the present, various methods have been attempted to treat Hunter syndrome,
such as bone marrow transplantation, enzyme supplementation, gene therapy, and the
like. The bone marrow transplantation has the disadvantages that although the symptom
is significantly improved, it is difficult to find a donor whose human leukocyte antigen
(HLA) matches with that of the patient and that the mortality rate before and after
surgery of the donor whose HLA does not match with that of the patent is high. The
gene therapy refers to a method in which a normal IDS gene is injected into the body
using a viral or non-viral vector such as an adenovirus or a retrovirus. However,
the gene therapy remains at an experimental level and is not yet clinically available.
[0004] Currently, the most widely used method is the enzyme replacement therapy (ERT) in
which a recombinant IDS enzyme is administered to a patient. Normally, the patient
visits the hospital once a week and is administered intravenously by professional
medical staff. It takes 3 to 4 hours or longer for a single administration.
[0005] Patients suffering from Hunter syndrome have great limitations in everyday life because
they have difficulties in catching objects or gait abnormality due to abnormalities
of the joint system, or they often have developmental disorders, cognitive disorders,
and behavior problems due to nervous system disorders. Therefore, the conventional
intravenous infusion therapy, which involves frequent visits to the hospital and long
treatment times, may lower the quality of life for the patients and their caregivers.
More importantly, there is a problem that the therapeutic effect is significantly
reduced due to the lowered compliance of the patents with the medication. Due to the
characteristics of the conventional treatment method of supplementing IDS by an intravenous
injection once a week, the concentration of IDS in the patient's body was the highest
immediately after the intravenous injection, but gradually decreases over time, thereby
increasing the concentration of GAG again in the body. An increase in the concentration
of GAG leads to severe aggravation of the symptoms. Further, given the high severity
and irreversibility of the symptoms of Hunter syndrome in general, if the patient
misses the appropriate treatment period, the resulting aggravation of the symptoms
can be very fatal and can greatly shorten the patient's life expectancy.
[0006] As discussed above, the intravenous administration of IDS in the conventional method
for treating Hunter syndrome has the problem that the therapeutic effect is greatly
restricted and the life expectancy of the patients can be shortened due to the lowered
compliance of the patients with the medication. Therefore, there is a pressing demand
for a new formulation and a treatment method to resolve the above-mentioned problem.
Disclosure of Invention
Technical Problem
[0007] An object of the present invention is to provide a formulation for treating Hunter
syndrome, which is capable of enhancing the therapeutic convenience of the patient.
[0008] Another object of the present invention is to provide a formulation for treating
Hunter syndrome, which can improve the patient's compliance with medication.
Solution to Problem
[0009]
- 1. A formulation for treating Hunter syndrome, comprising a first composition for
an intravenous injection and a second composition for a subcutaneous injection.
- 2. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the first composition is intravenously injected once every two months to twice a month,
and the second composition is subcutaneously injected 1 to 7 times a week.
- 3. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the first composition is intravenously injected once a month, and the second composition
is subcutaneously injected once a week.
- 4. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the first composition is injected at the first week of the month, and the second composition
is injected 1 to 7 times per week from the next week to the last week.
- 5. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the first composition is injected at the first week of the two months, and the second
composition is injected 1 to 7 times per week from the next week to the last week
of the two months.
- 6. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the first composition comprises iduronate-2-sulfatase consisting of at least one of
the amino acid sequences of SEQ ID NOS: 1 and 2.
- 7. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the second composition comprises iduronate-2-sulfatase consisting of at least one
of the amino acid sequences of SEQ ID NOS: 1 and 2.
- 8. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the first composition is injected at an effective dose of 0.05 mg/kg to 20 mg/kg per
week.
- 9. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the first composition is injected at an effective dose of 0.1 mg/kg to 5 mg/kg per
week.
- 10. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the second composition is injected at an effective dose of 0.1 mg/kg to 40 mg/kg per
week.
- 11. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the second composition is injected at an effective dose of 0.2 mg/kg to 10 mg/kg per
week.
- 12. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the second composition comprises at least one buffer selected from the group consisting
of sodium phosphate and L-histidine.
- 13. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the second composition comprises at least one stabilizer selected from the group consisting
of Polysorbate 20 and arginine.
- 14. The formulation for treating Hunter syndrome according to Item 1 above, wherein
the second composition comprises an absorption enhancer, which is hyaluronidase.
- 15. A method for treating Hunter syndrome, comprising intravenously injecting a first
composition and subcutaneously injecting a second composition.
- 16. The method for treating Hunter syndrome according to Item 15 above, wherein the
first composition is intravenously injected once every two months to twice a month,
and the second composition is subcutaneously injected 1 to 7 times a week.
- 17. The method for treating Hunter syndrome according to Item 15 above, wherein the
first composition is intravenously injected once a month, and the second composition
is subcutaneously injected once a week.
- 18. The method for treating Hunter syndrome according to Item 15 above, wherein the
first composition is injected at the first week of the month, and the second composition
is injected 1 to 7 times per week from the next week to the last week.
- 19. The method for treating Hunter syndrome according to Item 15 above, wherein the
first composition is injected at the first week of the two months, and the second
composition is injected 1 to 7 times per week from the next week to the last week
of the two months.
- 20. The method for treating Hunter syndrome according to Item 15 above, wherein the
first composition comprises iduronate-2-sulfatase consisting of at least one of the
amino acid sequences of SEQ ID NOS: 1 and 2.
- 21. The method for treating Hunter syndrome according to Item 15 above, wherein the
second composition comprises iduronate-2-sulfatase consisting of at least one of the
amino acid sequences of SEQ ID NOS: 1 and 2.
- 22. The method for treating Hunter syndrome according to Item 15 above, wherein the
first composition is injected at an effective dose of 0.05 mg/kg to 20 mg/kg per week.
- 23. The method for treating Hunter syndrome according to Item 15 above, wherein the
first composition is injected at an effective dose of 0.1 mg/kg to 5 mg/kg per week.
- 24. The method for treating Hunter syndrome according to Item 15 above, wherein the
second composition is injected at an effective dose of 0.1 mg/kg to 40 mg/kg per week.
- 25. The method for treating Hunter syndrome according to Item 15 above, wherein the
second composition is injected at an effective dose of 0.2 mg/kg to 10 mg/kg per week.
- 26. A formulation for treating Hunter syndrome, comprising a therapeutic composition
subcutaneously administered to a patient at a dose of 0.001 mL/hour to 100 mL/hour.
Advantageous Effects of Invention
[0010] The therapeutic formulation and the therapeutic method of the present invention exhibit
equivalent or better drug efficacy as compared with IV administration once a week.
[0011] The therapeutic formulation and the therapeutic method of the present invention exhibit
equivalent or better drug efficacy as compared with the conventional IV administration
once a week while reducing the number of visits of the patients suffering from Hunter
syndrome to the hospital to twice a month or less.
[0012] The therapeutic formulation and the therapeutic method of the present invention can
reduce the number of intravenous injections to the patients suffering from Hunter
syndrome.
[0013] The therapeutic formulation and the therapeutic method of the present invention can
improve the therapeutic convenience of the patients suffering from Hunter syndrome,
who have difficulties in visiting the hospital.
[0014] The therapeutic formulation and the therapeutic method of the present invention improve
the medication compliance of the patients suffering from Hunter syndrome with the
medicine, thereby improving the therapeutic effect.
[0015] The therapeutic formulation and the therapeutic method of the present invention are
suitable for effectively injecting iduronate-2-sulfatase consisting of a predetermined
amino acid sequence.
[0016] The present invention can provide a formulation for treating Hunter syndrome, which
improves its in vivo stability by comprising a buffer such as sodium phosphate and
L-histidine in the second composition.
[0017] The present invention can provide a formulation for treating Hunter syndrome, which
improves its storage and handling stability by comprising a stabilizer such as polysorbate
20 and arginine in the second composition.
[0018] The present invention can provide a formulation for treating Hunter syndrome, which
has an improved absorption rate in the body by comprising an absorption enhancer such
as hyaluronidase in the second composition.
Brief Description of the Drawings
[0019]
Fig. 1 shows the serum concentration of IDS with a single IV or SC administration
of the formulation for treating Hunter syndrome in Test Example 1.
Fig. 2 shows the concentration of GAG in the urine sample with a single IV or SC administration
of the formulation for treating Hunter syndrome in Test Example 2.
Fig. 3 shows the concentration of GAG in the spleen sample with a single IV or SC
administration of the formulation for treating Hunter syndrome in Test Example 2.
Fig. 4 shows the concentration of GAG in the heart sample with a single IV or SC administration
of the formulation for treating Hunter syndrome in Test Example 2.
Fig. 5 shows the concentration of GAG in the kidney sample with a single IV or SC
administration of the formulation for treating Hunter syndrome in Test Example 2.
Fig. 6 shows the concentration of GAG in the liver sample with a single IV or SC administration
of the formulation for treating Hunter syndrome in Test Example 2.
Fig. 7 shows the concentration of GAG in the lung sample with a single IV or SC administration
of the formulation for treating Hunter syndrome in Test Example 2.
Fig. 8 shows the concentration of GAG in the brain sample with a single IV or SC administration
of the formulation for treating Hunter syndrome in Test Example 2.
Fig. 9 shows the concentration of GAG in the kidney sample with repeated and combined
IV and SC administrations of the formulation for treating Hunter syndrome in Test
Example 4.
Fig. 10 shows the concentration of GAG in the liver sample with repeated and combined
IV and SC administrations of the formulation for treating Hunter syndrome in Test
Example 4.
Fig. 11 shows the concentration of GAG in the spleen sample with repeated and combined
IV and SC administrations of the formulation for treating Hunter syndrome in Test
Example 4.
Fig. 12 shows the concentration of GAG in the urine sample with repeated and combined
IV and SC administrations of the formulation for treating Hunter syndrome in Test
Example 4.
Mode for Carrying out the Invention
[0020] The present invention relates to a formulation for treating Hunter syndrome and more
specifically to a formulation for treating Hunter syndrome, which comprises a first
composition for an intravenous injection and a second composition for a subcutaneous
injection and is capable of exhibiting equivalent or better drug efficacy as compared
with the conventional IV administration once a week while reducing the number of visits
of the patients suffering from Hunter syndrome to the hospital to twice a month or
less; improving the compliance of the patients suffering from Hunter syndrome with
the medicine as compared with the conventional therapeutic formulation and method;
and enhancing the welfare and convenience of the patients suffering from Hunter syndrome.
The method of combined IV/SC administrations according to the present invention replaces
a certain number of IV administrations in the conventional method of IV administration
once a week, which requires that the patients visit the hospital every week, with
SC administrations that can be made by the patents by themselves at home. The method
of the present invention requires that the patents visit the hospital less often than
the conventional method of IV administration once a week, while exhibiting equivalent
or better drug efficacy as compared with the conventional IV administration once a
week. Thus, the present invention starkly contradicts the conventional common idea
that the therapeutic effect of SC administrations will not be superior to the therapeutic
effect of IV administrations. Accordingly, the present invention relates to a formulation
and a method, which drastically improve the therapeutic effect on Hunter syndrome
as compared with the conventional therapy.
[0021] The present invention relates to a formulation for treating Hunter syndrome, which
comprises a first composition for an intravenous injection and a second composition
for a subcutaneous injection; and a method for treating Hunter syndrome, which comprises
combined administrations of intravenously injecting a first composition and subcutaneously
injecting a second composition.
[0022] Hereinafter, the present invention will be described in detail.
[0023] The formulation for treating Hunter syndrome according to the present invention comprises
a first composition for an intravenous injection and a second composition for a subcutaneous
injection.
[0024] The first composition is a composition for an intravenous injection. A composition
for an intravenous injection refers to a sterile composition, which allows a drug
in liquid phase to be injected directly into the vein to act. A composition for injection
covers all compositions that can be used in the preparation of a customary injection
and includes, but is not limited to, aqueous injections, non-aqueous injections, suspension
injections, and freeze-dried injections.
[0025] The first composition is intravenously injected once every two months to twice a
month.
[0026] The administration once a month means that the interval between the previous intravenous
injection and the next intravenous injection is one month or so. For example, the
interval between intravenous injections may be 25 days, 26 days, 27 days, 28 days,
29 days, 30 days, 31 days, 32 days, or 33 days, depending on the conditions of the
patient or those of the treatment.
[0027] The first composition comprises an active ingredient that is effective in treating
Hunter syndrome.
[0028] Iduronate-2-sulfatase (IDS or I2S) may be used as an active ingredient in the first
composition.
[0029] IDS in the present invention comprises, for example, a protein consisting of the
amino acid sequence of SEQ ID NO: 1 or 2. The protein consisting of the amino acid
sequence of SEQ ID NO: 2 has the 59
th cysteine (Cys) of the protein consisting of the amino acid sequence of SEQ ID NO:
1 substituted by formylglycine (FGly).
[0030] IDS comprises proteins in which some amino acids have been inserted, deleted, substituted,
or the like, in the protein consisting of the amino acid sequence of SEQ ID NO: 1
or 2, as long as the therapeutic activity for treating Hunter syndrome is maintained.
[0031] IDS may be a protein derived from an animal such as a human or may be a recombinant
protein.
[0032] IDS may be a mixture of two or more proteins. For example, IDS may be a mixture of
a protein consisting of the amino acid sequence of SEQ ID NO: 1 and a protein consisting
of the amino acid sequence of SEQ ID NO: 2. In addition, IDS may be, for example,
a mixture of 35% (by mole) or less of a protein consisting of the amino acid sequence
of SEQ ID NO: 1 and 65% or more, 70% or more, 75% or more, or 80% (by mole in each
occurrence) or more of a protein consisting of the amino acid sequence of SEQ ID NO:
2.
[0033] If the protein consisting of the amino acid sequence of SEQ ID NO: 2 is contained
in an amount of 65% (by mole) or more, the therapeutic effect on Hunter syndrome is
improved.
[0034] The protein consisting of the amino acid sequence of SEQ ID NO: 1 or 2 may have mannose-6-phosphate
(M6P) in an amount of 2.0 to 4.0 moles, preferably 2.3 to 3.5 moles, more preferably
2.5 to 3.0 moles, per 1 mole of the protein. Since M6P contributes to the uptake of
IDS into the cells and the targeting of lysosomes, it is possible to effectively decompose
glycosaminoglycan accumulated in the lysosomes if the content of M6P is high.
[0035] The first composition may comprise various inactive ingredients required for the
intravenous injection composition other than the active ingredient (i.e., effective
ingredient).
[0036] For example, the first composition may further comprise buffers (such as sodium phosphate
and L-histidine), carbohydrates (such as glucose, mannose, sucrose, and dextran),
stabilizers (such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid, Polysorbate
20, and arginine), antioxidants, bacteriostatic agents, chelating agents (such as
EDTA and glutathione), adjuvants (such as aluminum hydroxide), suspensions, thickeners,
and/or preservatives (such as benzalkonium chloride, methyl- or propyl-paraben, and
chlorobutanol). In addition, it may further comprise various antibacterial agents
and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, and thimerosal.
[0037] The first composition may comprise a liquid suitable for an intravenous injection.
This liquid may be, but is not limited to, solvents or dispersion media comprising
water, ethanol, polyols (such as glycerol, propylene glycol, and liquid polyethylene
glycol), mixtures thereof and/or vegetable oils. More preferably, an isotonic solution
such as saline, a Hanks' solution, a ringer solution, PBS (phosphate buffered saline)
containing triethanolamine, or sterilized water for injection, 10% ethanol, 40% propylene
glycol, and 5% dextrose may be used. In addition, Other liquids suitable for intravenous
administration may also be referenced to
Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA,
1995.
[0038] The second composition is a composition for a subcutaneous injection. A composition
for a subcutaneous injection refers to a sterile composition, which allows a drug
in liquid phase to be injected into the loose connective tissues below the dermis
and absorbed through the capillary blood vessels to act.
[0039] The second composition is subcutaneously injected once to seven times a week.
[0040] The administration once to seven times a week means that the administration is performed
at one time to seven times a week at regular intervals. For example, administration
twice a week may be conducted on Monday and Thursday, on Tuesday and Friday, on Wednesday
and Saturday, on Thursday and Sunday, on Friday and Monday, on Saturday and Tuesday,
or on Sunday and Wednesday. Further, for example, administration three times a week
may be performed on Monday, Wednesday, and Friday; on Tuesday, Thursday, and Saturday;
on Wednesday, Friday, and Sunday; on Thursday, Saturday, and Monday; on Friday, Sunday,
and Tuesday; or on Saturday, Monday, and Wednesday.
[0041] The second composition may be prepared basically by comprising components the same
as, or similar to, those of the first composition (for example, the active ingredient
(i.e., effective ingredient) and the inactive ingredients). These components may be
changed for use as needed.
[0042] The second composition may comprise a liquid suitable for a subcutaneous injection.
This liquid may be the same as, or similar to, that contained in the first composition
and may be changed for use as needed.
[0043] The second composition may comprise sodium phosphate and L-histidine (20 mM, pH 6.0)
as a buffer. The addition of a buffer can contribute to stabilization of the active
pharmaceutical ingredient (API) in vivo even at a transient pH change.
[0044] The second composition may comprise Polysorbate 20 and arginine as a stabilizer.
The concentration of Polysorbate 20 may be about 0.05 to 0.22 mg/mL.
[0045] Arginine may serve as a stabilizer and a solubilizer in addition to a buffer.
[0046] The second composition may comprise hyaluronidase as an absorption enhancer, which
can enhance the absorption rate of the active ingredient in the body.
[0047] The second composition may comprise saline. The concentration of sodium chloride
may be 2% or less, and the acidity (pH) may be from 3 to 8.
[0048] The order of injection of the first and second compositions may be such that the
first composition is injected first, followed by injection of the second composition;
or that the second composition is injected first, followed by injection of the first
composition. For example, an intravenous injection may be administered at week 1,
and subcutaneous injections may be administered at weeks 2 to 4; a subcutaneous injections
may be administered at week 1, an intravenous injection may be administered at week
2, and subcutaneous injections may be administered at weeks 3 and 4; subcutaneous
injections may be administered at weeks 1 and 2, an intravenous injection may be administered
at week 3, and a subcutaneous injection may be administered at week 4; or subcutaneous
injections may be administered at weeks 1-3 and an intravenous injection may be administered
at week 4.
[0049] The first composition may be administered at various effective doses (i.e., the weight
of the active ingredient administered per 1 kilogram of body weight to be treated
in the unit of mg/kg or mpk) depending on the severity of the disease. Typically,
it may be administered at an effect dose of 0.05 mg/kg to 20 mg/kg per week (for example,
from 0.1 mg/kg to 5 mg/kg per week, from 0.5 mg/kg to 2 mg/kg per week, or from 0.5
mg/kg to 1 mg/kg per week). According to a more specific example, it may be administered
at an effective dose of 0.5 mg/kg per week.
[0050] The second composition may be administered at various effective doses depending on
the severity of the disease. Typically, it may be administered at an effect dose of
0.1 mg/kg to 40 mg/kg per week (for example, from 0.2 mg/kg to 20 mg/kg per week,
from 0.5 mg/kg to 10 mg/kg per week, or from 0.5 mg/kg to 5 mg/kg per week). According
to a more specific example, it may be administered at an effective dose of 1 mg/kg
to 5 mg/kg per week (for example, from 2.5 mg/kg to 5 mg/kg per week). Also, the single
dose volume of the second composition may be 2 mL/site or less (for example, 1 mL/site
or less), and the concentration of IDS may be 1 to 300 mg/mL.
[0051] The weekly dose of the second composition may be such that the concentration of IDS
in the patient's serum is 10 to 10,000 ng/mL within 24 hours after administration;
that the average maximum serum concentration (C
max) is 1.5 µg/mL or more; and that the area under the concentration-time curve (AUC)
is 200 to 1,000 min*µg/mL or more.
[0052] The administration of the second composition may be such that IDS is delivered to
tissues selected from the group consisting of muscle, skin, liver, kidney, spleen,
joint, bone, lung, airway, tongue, upper respiratory tract, eye, ear, connective tissue,
and heart; or that the concentration of IDS in the above-mentioned tissues may be
increased. The administration of the second composition may cause the activity of
IDS in the tissues to increase by at least 1 to 10 folds or more of the control group;
that the increased activity is 10 to 600 nmole/hr/mg or more; and that the increased
activity is 10 to 95% or more of the normal IDS activity.
[0053] The administration of the second composition may be such that the concentration of
GAG in the serum, plasma, urine, and above-mentioned tissues is reduced by 10 to 100%
of the difference in the concentration of GAG between the control group (i.e., control
group before administration or untreated) and the normal group (i.e., normal tissues
that are not affected by Hunter syndrome); and that the size of the liver or spleen
may be reduced by 10 to 100% of the difference in size between the control group (i.e.,
control group before administration or untreated) and the normal group (i.e., normal
tissues that are not affected by Hunter syndrome).
[0054] The administration of the second composition may be such that the result in a 6-minute
gait test is improved by 10 to 250 meters or more over the control group (i.e., control
group before administration or untreated) and by 10 to 1,000% or more over the control
group (i.e., control group before administration or untreated).
[0055] Hereinafter, the present invention will be described in more detail with reference
to the following examples and test examples. However, these examples and test examples
are set forth to illustrate the present invention in detail, and the scope of the
present invention is not limited thereto.
[0056] In the following test examples, IV or I.V stands for intravenous injection. SC or
S.C stands for subcutaneous injection. PK refers to pharmacokinetics. PD stands for
pharmacodynamics. Also, WT stands for wild-type, and KO stands for IDS knock-out.
Test Example 1: Pharmacokinetic study on the treatment of Hunter syndrome with a single
IV or SC administration
[0057] An IV composition and an SC composition were administered to mice to measure the
serum concentration of IDS, and such pharmacokinetic analysis as AUC and bioavailability
were carried out. Here, 6 to 8-week-old male mice were used. The experiment design
is summarized in Table 1.
[Table 1]
Pharmacokinetic experiment design with single administration |
Group (start age) |
Subgroup (3/time point) |
Route |
Dosing Regimen (mg/kg) |
PK time points |
WT male |
N=18 |
I.V |
5 |
5 min, 30 min, 1 hr, 3 hr, 6 hr, 8 hr |
(n=72) (6-8-week-old) |
N=18 |
S.C |
5 |
1 hr, 2 hr, 8 hr, 12 hr, 16 hr, 24 hr |
N=18 |
S.C |
10 |
1 hr, 2 hr, 8 hr, 12 hr, 16 hr, 24 hr |
N=18 |
S.C |
20 |
1 hr, 2 hr, 8 hr, 12 hr, 16 hr, 24 hr |
[0058] Seventy-two normal mice were divided into four groups. One group was administered
with an intravenous injection of 5 mg/kg corresponding to 10 times the clinical dose
(0.5 mg/kg) of an intravenous injection (IV), and the other three groups were administered
with a subcutaneous injection of 5, 10, and 20 mg/kg, respectively. The serum concentration
of IDS was analyzed by ELISA. The results are shown in Fig. 1. Phoenix ™ WinNonlin®
(ver 6.4, Pharsight)/NCA (non-compartmental analysis) was used for the PK analysis.
The results of pharmacokinetic analysis are summarized in Table 2.
[Table 2]
Results of pharmacokinetic analysis with single administration |
PK parameters |
Intravenous injection |
Subcutaneous injection |
5 mg/kg |
5 mg/kg |
10 mg/kg |
20 mg/kg |
Cmax (µg/mL) |
66.8 ± 1.57 |
1.41 ± 0.30 |
2.90 ± 0.687 |
8.48 ± 1.27 |
Cmax /D (kg·µg/mL/mg) |
- |
0.281 ± 0.06 |
0.290 ± 0.0687 |
0.424 ± 0.0636 |
C0 (µg/mL) |
82.4 ± 1.28 |
- |
- |
- |
Clast (µg/mL) |
0.176 ± 0.0187 |
0.116 ± 0.002 |
0.113 ± 0.0474 |
0.393 ± 0.0459 |
Tmax (hr) |
- |
1.00 ± 0.00 |
1.00 ± 0.00 |
1.00 ± 0.00 |
T1/2 (hr, terminal phase) |
1.80 ± 0.218 |
6.89 ± 0.952 |
5.25 ± 0.501 |
5.60 ± 0.567 |
AUCINF (hr·µg/mL) |
48.2 ± 3.61 |
10.6 ± 0.958 |
19.2 ± 4.02 |
53.9 ± 0.444 |
AUCINF/D (hr·kg· µg/mL/mg) |
- |
2.12 ± 0.192 |
1.92 ± 0.402 |
2.69 ± 0.0222 |
AUC%_Extrap (%) |
0.969 ± 0.279 |
11.0 ± 2.24 |
4.37 ± 1.39 |
5.94 ± 1.27 |
Cl (mL/hr/kg) |
104.1 ± 7.97 |
- |
- |
- |
Vss (mL/kg) |
83.3 ± 3.99 |
- |
- |
- |
MRTINF (hr) |
0.801 ± 0.0235 |
10.0 ± 1.16 |
7.29 ± 0.362 |
7.52 ± 0.765 |
BA (%) |
- |
22.0 |
19.9 |
27.9 |
[0059] As shown in the table above, IDS reached the maximum serum concentration (C
max, µg/mL) at a rapid rate within 1 hour in the case of SC administration. The half-life
(T
1/2, hr) was 1.80 hr in the terminal phase in the case of IV administration, whereas
the half-life was increased to 5.25 to 6.89 hr in the case of SC administration. The
mean retention time (MRT, hr) was 0.801 hr in the group administered with an IV injection
and 7.29 to 10.0 hr in the group administered with an SC injection. The concentration
(C
0, µg/mL) immediately after an IV administration was 82.4 µg/mL, which confirmed that
most of the dose (5 mg/kg) was recovered. The C
max and AUC
INF values, which show the body's exposure to drugs upon an SC administration, increased
dose-dependently. However, the increase was slightly greater in the group administered
with 20 mg/kg than in the groups administered with 5 and 10 mg/kg. The AUC% _Extrap
value was 0.969 to 11.0% in all treatment groups.
[0060] The bioavailability (BA, %) values were 22.0%, 19.9%, and 27.9% in the groups SC
administered with 5, 10, and 20 mg/kg (with reference to AUC when 5 mg/kg was SC administered
and with an assumption of linear PK), respectively. Overall, the BA values were approximately
20% when compared with the case of an IV administration. Therefore, it is expected
that an SC administration at a dose of approximately 5 times (2.5 mg/kg) the IV dose
will show a similar therapeutic effect to that of an IV administration.
Test Example 2: Pharmacodynamic study on the treatment of Hunter syndrome with a single
IV or SC administration
[0061] In consideration of the pharmacokinetic analysis (i.e., the BA value of an SC injection
was about 20% of that of an IV injection), the effective dose of an SC injection was
set to 2.5 mg/kg. After a single IV or SC injection, the effect thereof on reduction
in the concentration of GAG was compared for 4 weeks. Urine samples were collected
on the last day of Week 2 (i.e., Day 14) and on the last day of Week 4 (i.e., Day
28) counting from the day of dosing (Day 0). Samples of tissues (liver, brain, heart,
kidney, spleen, and lung) were also collected on the last day of Week 4 (i.e., Day
28) for the measurement of GAG concentrations. A urine samples was collected once
for three days prior to the drug administration for comparison. The pharmacodynamic
experiment design is summarized in Table 3.
[Table 3]
Pharmacodynamic experiment design with single administration |
Group (start age) |
Test item |
Subgroup |
Route |
Dosing Regimen (mg/kg) |
WT (n=6) |
Vehicle |
N=6 |
S.C |
- |
KO (n=30) (6-8-week-old) |
Vehicle |
N=6 |
S.C |
- |
Formulation for treating Hunter syndrome |
N=6 |
S.C |
0.5 |
N=6 |
S.C |
2.5 |
N=6 |
S.C |
12.5 |
Formulation for treating Hunter syndrome |
N=6 |
I.V |
0.5 |
[0062] The results of GAG concentration analysis of the urine samples according to the experiment
design shown in Table 3 are summarized in Table 4 and Fig. 2.
[Table 4]
Results of GAG concentration analysis of the urine samples with single administration
(Day 28) |
Group |
GAG (µg/µg of creatinine) |
P-value |
Summary |
WT (n=6) |
0.577 ± 0.155 |
<0.001 |
*** |
KO (n=6) |
1.29 ± 0.0958 |
- |
- |
SC |
0.5 mg/kg (n=6) |
1.07 ± 0.0975 |
<0.05 |
* |
2.5 mg/kg (n=6) |
0.867 ± 0.101 |
<0.001 |
*** |
12.5 mg/kg (n=6) |
0.815 ± 0.113 |
<0.001 |
*** |
IV |
0.5 mg/kg (n=6) |
0.923 ± 0.136 |
<0.001 |
*** |
The GAG concentration values were expressed as Mean ± SEM.
*: p<0.05, **: p<0.01, ***: p<0.001 vs KO.
One-way ANOVA and Dunnett's Multiple Comparison Test (GraphPad Prism) |
[0063] The results of GAG concentration analysis of the spleen samples according to the
experiment design shown in Table 3 are summarized in Table 5 and Fig. 3.
[Table 5]
Results of GAG concentration analysis of the spleen samples with single administration
(Day 28) |
Group |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=6) |
1.41 ± 0.121 |
<0.001 |
*** |
KO (n=6) |
8.55 ± 0.619 |
- |
- |
sc |
0.5 mg/kg (n=6) |
5.88 ± 0.329 |
<0.001 |
*** |
2.5 mg/kg (n=6) |
5.89 ± 0.524 |
<0.001 |
*** |
|
12.5 mg/kg (n=6) |
4.03 ± 0.362 |
<0.001 |
*** |
IV |
0.5 mg/kg (n=6) |
4.80 ± 0.470 |
<0.001 |
*** |
The GAG concentration values were expressed as Mean ± SEM.
*: p<0.05, **: p<0.01, ***: p<0.001 vs KO.
One-way ANOVA and Dunnett's Multiple Comparison Test (GraphPad Prism) |
[0064] The results of GAG concentration analysis of the heart samples according to the experiment
design shown in Table 3 are summarized in Table 6 and Fig. 4.
[Table 6]
Results of GAG concentration analysis of the heart samples with single administration
(Day 28) |
Group |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=6) |
0.390 ± 0.0594 |
<0.001 |
### |
KO (n=6) |
11.1 ± 0.560 |
- |
- |
SC |
0.5 mg/kg (n=6) |
10.7 ± 0.799 |
>0.05 |
- |
2.5 mg/kg (n=6) |
6.75 ± 0.405 |
>0.05 |
- |
12.5 mg/kg (n=6) |
3.49 ± 0.212 |
<0.01 |
## |
IV |
0.5 mg/kg (n=6) |
5.96 ± 0.0613 |
>0.05 |
- |
The GAG concentration values were expressed as Mean ± SEM.
#: p<0.05, ##: p<0.01, ###: p<0.001 vs KO.
Kruskal-Wallis Test and Dunn's Multiple Comparison Test (GraphPad Prism) |
[0065] The results of GAG concentration analysis of the kidney samples according to the
experiment design shown in Table 3 are summarized in Table 7 and Fig. 5.
[Table 7]
Results of GAG concentration analysis of the kidney samples with single administration
(Day 28) |
Group |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=6) |
0.639 ± 0.0593 |
<0.01 |
## |
KO (n=6) |
25.0 ± 0.957 |
- |
- |
SC |
0.5 mg/kg (n=6) |
25.9 ± 0.645 |
>0.05 |
- |
2.5 mg/kg (n=6) |
23.8 ± 0.821 |
>0.05 |
- |
12.5 mg/kg (n=6) |
16.4 ± 0.740 |
>0.05 |
- |
IV |
0.5 mg/kg (n=6) |
23.5 ± 1.34 |
>0.05 |
- |
The GAG concentration values were expressed as Mean ± SEM.
#: p<0.05, ##: p<0.01, ###: p<0.001 vs KO.
Kruskal-Wallis Test and Dunn's Multiple Comparison Test (GraphPad Prism) |
[0066] The results of GAG concentration analysis of the liver samples according to the experiment
design shown in Table 3 are summarized in Table 8 and Fig. 6.
[Table 8]
Results of GAG concentration analysis of the liver samples with single administration
(Day 28) |
Group |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=6) |
0.676 ± 0.0143 |
<0.001 |
### |
KO (n=6) |
50.5 ± 1.19 |
- |
- |
SC |
0.5 mg/kg (n=6) |
23.7 ± 0.159 |
>0.05 |
- |
2.5 mg/kg (n=6) |
18.6 ± 0.815 |
>0.05 |
- |
12.5 mg/kg (n=6) |
10.3 ± 0.799 |
<0.01 |
## |
IV |
0.5 mg/kg (n=6) |
16.0 ± 0.762 |
>0.05 |
- |
The GAG concentration values were expressed as Mean ± SEM.
#: p<0.05, ##: p<0.01, ###: p<0.001 vs KO.
Kruskal-Wallis Test and Dunn's Multiple Comparison Test (GraphPad Prism) |
[0067] The results of GAG concentration analysis of the lung samples according to the experiment
design shown in Table 3 are summarized in Table 9 and Fig. 7.
[Table 9]
Results of GAG concentration analysis of the lung samples with single administration
(Day 28) |
Group |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=6) |
0.992 ± 0.0951 |
<0.001 |
### |
KO (n=6) |
34.4 ± 2.43 |
- |
- |
SC |
0.5 mg/kg (n=6) |
31.3 ± 2.42 |
>0.05 |
- |
2.5 mg/kg (n=6) |
28.8 ± 2.10 |
>0.05 |
- |
12.5 mg/kg (n=6) |
19.2 ± 0.955 |
<0.05 |
# |
IV |
0.5 mg/kg (n=6) |
29.1 ± 2.11 |
>0.05 |
- |
The GAG concentration values were expressed as Mean ± SEM.
#: p<0.05, ##: p<0.01, ###: p<0.001 vs KO.
Kruskal-Wallis Test and Dunn's Multiple Comparison Test (GraphPad Prism) |
[0068] The results of GAG concentration analysis of the brain samples according to the experiment
design shown in Table 3 are summarized in Table 10 and Fig. 8.
[Table 10]
Results of GAG concentration analysis of the brain samples with single administration
(Day 28) |
Group |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=6) |
0.558 ± 0.0369 |
<0.001 |
*** |
KO (n=6) |
1.27 ± 0.0651 |
- |
- |
sc |
0.5 mg/kg (n=6) |
1.27 ± 0.0868 |
>0.05 |
- |
2.5 mg/kg (n=6) |
1.19 ± 0.0602 |
>0.05 |
- |
12.5 mg/kg (n=6) |
1.16 ± 0.0756 |
>0.05 |
- |
IV |
0.5 mg/kg (n=6) |
1.12 ± 0.0798 |
>0.05 |
- |
The GAG concentration values were expressed as Mean ± SEM.
*: p<0.05, **: p<0.01, ***: p<0.001 vs KO.
One-way ANOVA and Dunnett's Multiple Comparison Test (GraphPad Prism) |
[0069] Taking all the results shown in Tables 4 to 10 into consideration, it can be seen
that the formulation for treating Hunter syndrome of the present invention generally
exhibits the effect of reducing GAG dose-dependently when administered by a single
subcutaneous (SC) injection. Further, when 2.5 mg/kg is administered by a subcutaneous
(SC) injection, the effect of reducing GAG is similar to that of the clinical dose
of 0.5 mg/kg of an intravenous (IV) injection.
Test Example 3: Determination of SC infusion rate of the formulation for treating
Hunter syndrome
[0070] A commercially marketed vial for an intravenous injection of IDS has a size of 3.0
mL, which contains a solution that comprises 6.0 mg of an IDS enzyme in a concentration
of 2.0 mg/mL. This medicine is for one-time use. The recommended dose to the patients
is 0.5 mg per 1 kg of the body weight, which is gradually administered to the patients
intravenously when the patient visits the hospital once a week. The amount corresponding
to the patient's body weight is diluted in 100 mL of 0.9% sodium chloride water for
injection, which is administered intravenously. The total dose should be gradually
administered over 1 to 3 hours or longer. The infusion time may be extended due to
any infusion related reactions. However, the infusion time should not exceed 8 hours.
The initial infusion rate should be 8 mL/hour for 15 minutes from the beginning of
infusion, and the infusion rate may then be increased by 8 mL/hour at 15-minute intervals
to allow the total dose to be administered within the expected time if no toxicity
appears. However, doctors and nurses are instructed that the infusion rate should
not exceed a maximum of 100 mL/hour.
[0071] For a subcutaneous injection, a relatively small volume is generally administered
at once. However, a drug may be administered in a continuous SC or SC infusion if
there is a limit that the drug is concentrated to reduce the total volume of the drug.
In the case where a drug originally developed for an intravenous injection is changed
for a subcutaneous injection as in the present invention, whether the drug is suitable
for a continuous SC or SC infusion should be clearly confirmed. According to the present
invention, a subcutaneous administration may be carried out at a volume of 2 mL/site
or less. According to a more specific example, the dosage may be 1 mL/site or less.
Test Example 4: Pharmacodynamic study on the treatment of Hunter syndrome with repeated
and combined IV and SC administrations
[0072] The active ingredient of Idursulfase beta according to one embodiment of the present
invention was used. The effective dose for IV was 0.5 mg/kg, and the effective dose
for SC was 0.5 mg/kg, 1 mg/kg, 2.5 mg/kg, or 5 mg/kg. Mice were subjected to repeated
IV administrations, repeated SC administrations, and repeated IV and SC administrations
in accordance with the experiment design as shown in Table 11 below. In the case where
the effective dose for IV and SC is 0 mg/kg in Table 11, it means a control group
in which only saline was administered.
[Table 11]
Mouse |
Substance |
Weekly dosing scheme |
Day 0 |
Day 7 |
Day 14 |
Day 21 |
Day 28 |
Day 35 |
Day 42 |
Day 49 |
WT |
Saline |
IV (0) |
SC (0) |
sc (0) |
SC (0) |
IV (0) |
SC (0) |
SC (0) |
SC (0) |
IDS KO |
Saline |
IV (0) |
SC (0) |
SC (0) |
SC (0) |
IV (0) |
SC (0) |
SC (0) |
SC (0) |
|
Idursulfase beta |
IV (0.5) |
SC (0.5) |
SC (0.5) |
SC (0.5) |
IV (0.5) |
SC (0.5) |
SC (0.5) |
SC (0.5) |
IV (0.5) |
SC (1) |
sc (1) |
SC (1) |
IV (0.5) |
SC (1) |
SC (1) |
SC (1) |
IV (0.5) |
sc (2.5) |
SC (2.5) |
SC (2.5) |
IV (0.5) |
SC (2.5) |
SC (2.5) |
SC (2.5) |
IV (0.5) |
SC (5) |
SC (5) |
SC (5) |
IV (0.5) |
SC (5) |
SC (5) |
SC (5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
SC (2.5) |
SC (2.5) |
SC (2.5) |
SC (2.5) |
SC (2.5) |
SC (2.5) |
SC (2.5) |
SC (2.5) |
Idursulfase |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
IV (0.5) |
* Day 0 refers to the first administration day, and Day 7, Day 14, Day 21, Day 28,
Day 35, Day 42, and Day 49 refer to the 7th, 14th, 21st, 28th, 35th, 42nd, and 49th days from the first administration day, respectively. |
* Urine samples were collected on Day -3, Day 14, Day 28, and Day 56; and Day -3 refers
to 3 days before the first administration day. |
* Tissue samples were collected on Day 56. |
* IV or SC stands for the administration route on the corresponding administration
day (IV: intravenous injection, SC: subcutaneous injection). |
* The number in parentheses after IV or SC indicates the effective dose of the formulation
for treating Hunter syndrome per administration in the unit of mg/kg. |
* Idursulfase beta is the active ingredient according to one embodiment of the present
invention. |
* Idursulfase was used as a control group. |
[0073] In accordance with the above experiment design, repeated IV administrations, repeated
SC administrations, and repeated IV and SC administrations were carried out in a total
of 8 times in each test at intervals of one week. Urine samples were collected on
three days before the first administration day, the 14
th day, 28
th day, and 56
th day from the first administration day, respectively. Tissue samples were collected
on the 56
th day (D 56). The effect of reducing GAG was then compared.
[0074] The results of GAG concentration analysis of the kidney, liver, and spleen samples
according to the experiment design shown in Table 11 are summarized in Tables 12 to
14 and Figs. 9 to 11.
[Table 12]
Results of GAG concentration analysis of the kidney samples with combined administrations
(Day 56) |
Mouse |
Substance |
Concentration |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=4) |
Saline |
- |
0.25±0.25 |
<0.001 |
*** |
IDS KO (n=4) |
Saline |
- |
20.65±0.30 |
- |
- |
Idursulfase beta |
0.5 IV → 0.5 SC |
13.66±1.40 |
<0.05 |
* |
0.5 IV → 1.0 SC |
13.98±0.94 |
<0.05 |
* |
0.5 IV → 2.5 SC |
10.86±1.22 |
<0.01 |
** |
0.5 IV → 5.0 SC |
6.61±2.66 |
<0.001 |
*** |
2.5 SC |
14.83±0.90 |
>0.05 |
- |
0.5 IV |
8.01±2.92 |
<0.001 |
*** |
Idursulfase |
0.5 IV |
10.47±1.92 |
<0.001 |
*** |
The GAG concentration values were expressed as Mean ± SEM. |
*: p<0.05, **: p<0.01, ***: p<0.001 vs KO. |
One-way ANOVA and Dunnett's Multiple Comparison Test (GraphPad Prism) |
[0075] As shown in Table 12 and Fig. 9, when the formulation for treating Hunter syndrome
was administered in combination of an effective IV dose of 0.5 mg/kg and an effective
SC dose of 2.5 mg/kg (see "0.5 IV → 2.5 SC" in Table 12 and Fig. 9) and when the formulation
for treating Hunter syndrome was administered in combination of an effective IV dose
of 0.5 mg/kg and an effective SC dose of 5.0 mg/kg (see "0.5 IV → 5.0 SC" in Table
12 and Fig. 9), the concentrations of GAG in the kidney were equivalent to, or lower
than, that of the repeated administrations of the formulation at an effective IV dose
of 0.5 mg/kg (see "0.5 IV" in Table 12 and Fig. 9).
[Table 13]
Results of GAG concentration analysis of the liver samples with combined administrations
(Day 56) |
Mouse |
Substance |
Concentration |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=4) |
Saline |
- |
0.94±0.09 |
<0.001 |
*** |
IDS KO (n=4) |
Saline |
- |
30.05±0.65 |
- |
- |
Idursulfase beta |
0.5 IV → 0.5 SC |
1.60±0.43 |
<0.001 |
*** |
0.5 IV → 1.0 SC |
1.31±0.38 |
<0.001 |
*** |
0.5 IV → 2.5 SC |
1.58±0.42 |
<0.001 |
*** |
0.5 IV→ 5.0 SC |
0.81±0.18 |
<0.001 |
*** |
2.5 SC |
2.75±0.66 |
<0.001 |
*** |
0.5 IV |
0.91±0.17 |
<0.001 |
*** |
Idursulfase |
0.5 IV |
1.07±0.21 |
<0.001 |
*** |
The GAG concentration values were expressed as Mean ± SEM. |
*: p<0.05, **: p<0.01, ***: p<0.001 vs KO. |
One-way ANOVA and Dunnett's Multiple Comparison Test (GraphPad Prism) |
[0076] As shown in Table 13 and Fig. 10, when the formulation for treating Hunter syndrome
was administered in combination of an effective IV dose of 0.5 mg/kg and an effective
SC dose of 0.5 mg/kg (see "0.5 IV → 0.5 SC" in Table 13 and Fig. 10), when the formulation
for treating Hunter syndrome was administered in combination of an effective IV dose
of 0.5 mg/kg and an effective SC dose of 1.0 mg/kg (see "0.5 IV → 1.0 SC" in Table
13 and Fig. 10), when the formulation for treating Hunter syndrome was administered
in combination of an effective IV dose of 0.5 mg/kg and an effective SC dose of 2.5
mg/kg (see "0.5 IV → 2.5 SC" in Table 13 and Fig. 10), and when the formulation for
treating Hunter syndrome was administered in combination of an effective IV dose of
0.5 mg/kg and an effective SC dose of 5.0 mg/kg (see "0.5 IV → 5.0 SC" in Table 13
and Fig. 10), the concentrations of GAG in the liver were equivalent to, or lower
than, that of the repeated administrations of the formulation at an effective IV dose
of 0.5 mg/kg (see "0.5 IV" in Table 13 and Fig. 10).
[Table 14]
Results of GAG concentration analysis of the spleen samples with combined administrations
(Day 56) |
Mouse |
Substance |
Concentration |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=4) |
Saline |
- |
0.00±0.00 |
<0.001 |
*** |
IDS KO (n=4) |
Saline |
- |
4.64±0.43 |
- |
- |
Idursulfase beta |
0.5 IV → 0.5 SC |
1.20±0.44 |
<0.001 |
*** |
0.5 IV → 1.0 SC |
0.77±0.27 |
<0.001 |
*** |
0.5 IV → 2.5 SC |
0.85±0.53 |
<0.001 |
*** |
0.5 IV → 5.0 SC |
0.45±0.27 |
<0.001 |
*** |
2.5 SC |
1.43±0.15 |
<0.001 |
*** |
0.5 IV |
0.68±0.31 |
<0.001 |
*** |
Idursulfase |
0.5 IV |
0.65±0.23 |
<0.001 |
*** |
The GAG concentration values were expressed as Mean ± SEM. |
*: p<0.05, **: p<0.01, ***: p<0.001 vs KO. |
One-way ANOVA and Dunnett's Multiple Comparison Test (GraphPad Prism) |
[0077] As shown in Table 14 and Fig. 11, when the formulation for treating Hunter syndrome
was administered in combination of an effective IV dose of 0.5 mg/kg and an effective
SC dose of 1.0 mg/kg (see "0.5 IV → 1.0 SC" in Table 14 and Fig. 11), when the formulation
for treating Hunter syndrome was administered in combination of an effective IV dose
of 0.5 mg/kg and an effective SC dose of 2.5 mg/kg (see "0.5 IV → 2.5 SC" in Table
14 and Fig. 11), and when the formulation for treating Hunter syndrome was administered
in combination of an effective IV dose of 0.5 mg/kg and an effective SC dose of 5.0
mg/kg (see "0.5 IV → 5.0 SC" in Table 14 and Fig. 11), the concentrations of GAG in
the spleen were equivalent to, or lower than, that of the repeated administrations
of the formulation at an effective IV dose of 0.5 mg/kg (see "0.5 IV" in Table 14
and Fig. 11).
[0078] In accordance with the experiment design shown in Table 11, urine samples were collected
on Day 0, Day 14, Day 28, and Day56. The results of GAG concentration analysis are
summarized in Table 15 and Fig. 12.
[Table 15]
Results of GAG concentration analysis of the urine samples with combined administrations
(Day 56) |
Mouse |
Substance |
Concentration |
GAG (µg/mg of protein) |
P-value |
Summary |
WT (n=4) |
Saline |
- |
0.14±0.02 |
<0.01 |
** |
IDS KO (n=4) |
Saline |
- |
0.49±0.04 |
- |
- |
Idursulfase beta |
0.5 IV → 0.5 SC |
0.27±0.06 |
>0.05 |
- |
0.5 IV → 1.0 SC |
0.26±0.07 |
<0.05 |
* |
0.5 IV → 2.5 SC |
0.31±0.06 |
>0.05 |
- |
0.5 IV → 5.0 SC |
0.24±0.02 |
<0.05 |
* |
2.5 SC |
0.39±0.09 |
>0.05 |
- |
0.5 IV |
0.32±0.01 |
>0.05 |
- |
Idursulfase |
0.5 IV |
0.25±0.03 |
<0.05 |
* |
The GAG concentration values were expressed as Mean ± SEM. |
*: p<0.05, **: p<0.01, ***: p<0.001 vs KO. |
One-way ANOVA and Dunnett's Multiple Comparison Test (GraphPad Prism) |
[0079] As shown in Table 15 and Fig. 12, when the formulation for treating Hunter syndrome
was administered in combination of an effective IV dose of 0.5 mg/kg and an effective
SC dose of 1.0 mg/kg (see "0.5 IV → 1.0 SC" in Table 15 and Fig. 12), when the formulation
for treating Hunter syndrome was administered in combination of an effective IV dose
of 0.5 mg/kg and an effective SC dose of 2.5 mg/kg (see "0.5 IV → 2.5 SC" in Table
15 and Fig. 12), and when the formulation for treating Hunter syndrome was administered
in combination of an effective IV dose of 0.5 mg/kg and an effective SC dose of 5.0
mg/kg (see "0.5 IV → 5.0 SC" in Table 15 and Fig. 12), the concentrations of GAG in
all of the urine samples collected on Day 14, Day 28, and Day 56 were equivalent to,
or lower than, that of the repeated administrations of the formulation at an effective
IV dose of 0.5 mg/kg (see "0.5 IV" in Table 15 and Fig. 12).
[0080] Taking into consideration all the results of experiments conducted in accordance
with the experiment design shown in Table 11, it can be seen that the method of combined
IV/SC administrations according to the present invention, particularly the combined
administrations of an effective IV dose of 0.5 mg/kg and an effective SC dose of 2.5
mg/kg to 5 mg/kg, had an effect in treating Hunter syndrome equivalent to, or better
than, that of the conventional IV administration (i.e., IV administrations at intervals
of 7 days).
[0081] Therefore, the formulation for combined IV/SC administrations and the method of combined
IV/SC administrations according to the present invention replace a certain number
of IV administrations in the conventional method of IV administration once a week,
which requires that the patients visit the hospital every week and takes 3 to 4 hours
or longer per administration, with SC administrations that can be made by the patents
by themselves at home. The formulation and the method of the present invention require
that the patents visit the hospital less often than the conventional method of IV
administration once a week, thereby eliminating the chances that the patients fail
to visit the hospital for treatment, which drastically improves the compliance of
the patients suffering from Hunter syndrome with the medicine. In addition, since
the formulation and the method of the present invention exhibit drug efficacy equivalent
to, or better than, that of the conventional IV administration once a week, it is
possible to treat Hunter syndrome more effectively as compared with the conventional
therapy.
Test Example 5: Active ingredient of the formulation for treating Hunter syndrome
[0082] The active ingredient that can be employed in the formulation and the method for
treating Hunter syndrome according to an embodiment of the present invention may comprise,
for example, SEQ ID NO: 1 and SEQ ID NO: 2.
Sequence Listing Free Text
1. A formulation for treating Hunter syndrome, comprising a first composition for an
intravenous injection and a second composition for a subcutaneous injection.
2. The formulation for treating Hunter syndrome according to claim 1, wherein the first
composition is intravenously injected once every two months to twice a month, and
the second composition is subcutaneously injected once to seven times a week.
3. The formulation for treating Hunter syndrome according to claim 1, wherein the first
composition is intravenously injected once a month, and the second composition is
subcutaneously injected once a week.
4. The formulation for treating Hunter syndrome according to claim 1, wherein the first
composition is injected at the first week of the month, and the second composition
is injected 1 to 7 times per week from the next week to the last week.
5. The formulation for treating Hunter syndrome according to claim 1, wherein the first
composition is injected at the first week of the two months, and the second composition
is injected 1 to 7 times per week from the next week to the last week of the two months.
6. The formulation for treating Hunter syndrome according to claim 1, wherein the first
composition comprises iduronate-2-sulfatase consisting of at least one of the amino
acid sequences of SEQ ID NOS: 1 and 2.
7. The formulation for treating Hunter syndrome according to claim 1, wherein the second
composition comprises iduronate-2-sulfatase consisting of at least one of the amino
acid sequences of SEQ ID NOS: 1 and 2.
8. The formulation for treating Hunter syndrome according to claim 1, wherein the first
composition is injected at an effective dose of 0.05 mg/kg to 20 mg/kg per week.
9. The formulation for treating Hunter syndrome according to claim 1, wherein the first
composition is injected at an effective dose of 0.1 mg/kg to 5 mg/kg per week.
10. The formulation for treating Hunter syndrome according to claim 1, wherein the second
composition is injected at an effective dose of 0.1 mg/kg to 40 mg/kg per week.
11. The formulation for treating Hunter syndrome according to claim 1, wherein the second
composition is injected at an effective dose of 0.2 mg/kg to 10 mg/kg per week.
12. The formulation for treating Hunter syndrome according to claim 1, wherein the second
composition comprises at least one buffer selected from the group consisting of sodium
phosphate and L-histidine.
13. The formulation for treating Hunter syndrome according to claim 1, wherein the second
composition comprises at least one stabilizer selected from the group consisting of
Polysorbate 20 and arginine.
14. The formulation for treating Hunter syndrome according to claim 1, wherein the second
composition comprises an absorption enhancer, which is hyaluronidase.
15. A method for treating Hunter syndrome, comprising intravenously injecting a first
composition and subcutaneously injecting a second composition.
16. The method for treating Hunter syndrome according to claim 15, wherein the first composition
is intravenously injected once every two months to twice a month, and the second composition
is subcutaneously injected 1 to 7 times a week.
17. The method for treating Hunter syndrome according to claim 15, wherein the first composition
is intravenously injected once a month, and the second composition is subcutaneously
injected once a week.
18. The method for treating Hunter syndrome according to claim 15, wherein the first composition
is injected at the first week of the month, and the second composition is injected
1 to 7 times per week from the next week to the last week.
19. The method for treating Hunter syndrome according to claim 15, wherein the first composition
is injected at the first week of the two months, and the second composition is injected
1 to 7 times per week from the next week to the last week of the two months.
20. The method for treating Hunter syndrome according to claim 15, wherein the first composition
comprises iduronate-2-sulfatase consisting of at least one of the amino acid sequences
of SEQ ID NOS: 1 and 2.
21. The method for treating Hunter syndrome according to claim 15, wherein the second
composition comprises iduronate-2-sulfatase consisting of at least one of the amino
acid sequences of SEQ ID NOS: 1 and 2.
22. The method for treating Hunter syndrome according to claim 15, wherein the first composition
is injected at an effective dose of 0.05 mg/kg to 20 mg/kg per week.
23. The method for treating Hunter syndrome according to claim 15, wherein the first composition
is injected at an effective dose of 0.1 mg/kg to 5 mg/kg per week.
24. The method for treating Hunter syndrome according to claim 15, wherein the second
composition is injected at an effective dose of 0.1 mg/kg to 40 mg/kg per week.
25. The method for treating Hunter syndrome according to claim 15, wherein the second
composition is injected at an effective dose of 0.2 mg/kg to 10 mg/kg per week.
26. A formulation for treating Hunter syndrome, comprising a therapeutic composition subcutaneously
administered to a patient at a dose of 0.001 mL/hour to 100 mL/hour.